Circulating miRNA profiling of plasma samples deriving from recently... | Download Scientific DiagramFigure - available from: DiabetologiaThis content is subject to copyright. Terms and conditions apply.DownloadView publicationCopy referenceCopy captionEmbed figureCirculating miRNA profiling of plasma samples deriving from recently diabetic and normoglycaemic NOD mice. (a) Hierarchical clustering heatmap showing colour-coded expression levels of miRNAs (blue, high expression; red, low expression) in plasma from recently diabetic and age-matched normoglycaemic NOD mice. (b) Volcano plot showing changes in miRNA levels between recently diabetic and normoglycaemic NOD mice. Levels in normoglycaemic mice were set at 0. Red lines indicate fold change and p value cut-offs. The larger data points indicate differentially expressed miRNAs. (c–i) Single-assay real-time RT-PCR validation of differentially expressed miRNAs was performed in the same plasma samples as those used for the miRNA screening. Values are expressed as 2−ΔCtdocumentclass[12pt]{minimal} usepackage{amsmath} usepackage{wasysym} usepackage{amsfonts} usepackage{amssymb} usepackage{amsbsy} usepackage{mathrsfs} usepackage{upgreek} setlength{oddsidemargin}{-69pt} begin{document}$$ {2}^{-Delta {mathrm{C}}_{mathrm{t}}} $$end{document}. Data are presented as mean ± SEM; *p ≤ 0.05, **p ≤ 0.01, Mann–Whitney U test. mmu, Mus musculus; NG, normoglycaemic; rno, Rattus norvegicusSource publication+2 miR-409-3p is reduced in plasma and islet immune infiltrates of NOD diabetic mice and is differentially expressed in people with type 1 diabetesArticleFull-text availableJan 2020 Giuliana Ventriglia Francesca Mancarella Guido Sebastiani[...] Francesco DottaAims/hypothesisMicroRNAs (miRNAs) are a novel class of potential biomarkers emerging in many diseases, including type 1 diabetes. Here, we aim to analyse a panel of circulating miRNAs in non-obese diabetic (NOD) mice and individuals with type 1 diabetes.MethodsWe adopted standardised methodologies for extracting miRNAs from small sample volumes...CiteDownload full-textCitations... Efficient biomarkers for the early identification and stratification of high-risk T1D subjects are still lacking (148). Of note, numerous studies explored the potential usefulness of whole plasma/serum miRNAs analysis to detect novel biomarkers of T1D (149)(150)(151)(152). However, the detection of miRNAs associated to different plasma/serum components could be the key to uncover novel and more specific biomarkers. ...Extracellular Vesicles in Immune System Regulation and Type 1 Diabetes: Cell-to-Cell Communication Mediators, Disease Biomarkers, and Promising Therapeutic ToolsArticleFull-text availableJun 2021 Giuseppina Emanuela Grieco Daniela Fignani Caterina Formichi Laura Nigi Francesco DottaExtracellular vesicles (EVs) are generated by cells of origin through complex molecular mechanisms and released into extracellular environment. Hence, the presence of EVs has been described in multiple biological fluids and in most cases their molecular cargo, which includes non-coding RNAs (ncRNA), messenger RNAs (mRNA), and proteins, has been reported to modulate distinct biological processes. EVs release and their molecular cargo have been demonstrated to be altered in multiple diseases, including autoimmune diseases. Notably, numerous evidence showed a relevant crosstalk between immune system and interacting cells through specific EVs release. The crosstalk between insulin-producing pancreatic β cells and immune system through EVs bidirectional trafficking has yet started to be deciphered, thus uncovering an intricate communication network underlying type 1 diabetes (T1D) pathogenesis. EVs can also be found in blood plasma or serum. Indeed, the assessment of circulating EVs cargo has been shown as a promising advance in the detection of reliable biomarkers of disease progression. Of note, multiple studies showed several specific cargo alterations of EVs collected from plasma/serum of subjects affected by autoimmune diseases, including T1D subjects. In this review, we discuss the recent literature reporting evidence of EVs role in autoimmune diseases, specifically focusing on the bidirectional crosstalk between pancreatic β cells and immune system in T1D and highlight the relevant promising role of circulating EVs as disease biomarkers.View... Previous studies have suggested that miR-137-3p may be associated with the production of insulin as well as the metabolism of lipids [21,22]. Ventriglia et al. demonstrated that the expression of miR-409-3p was downregulated in the plasma samples of diabetic mice [23]. Moreover, with the development of diabetes, there is a gradual decrease in the miR-409-3p levels in the plasma. ...MicroRNA expression profile and identification of novel microRNA biomarkers for metabolic syndromeArticleFull-text availableJan 2021 Guanzhi LiuYutian LeiSen LuoZhuo HuangXin HuangThe lack of efficient biomarkers is the main reason for the inaccurate early diagnosis and poor treatment outcomes of patients with metabolic syndrome (MetS). The current study aimed to identify several novel microRNA (miRNA) biomarkers for metabolic syndrome via high-throughput sequencing and comprehensive bioinformatics analysis. Through high-throughput sequencing and differentially expressed miRNA (DEM) analysis, we first identified two upregulated and 36 downregulated DEMs in the plasma samples of patients with MetS compared to the healthy volunteers. Additionally, we also predicted 379 potential target genes and subsequently carried out enrichment analysis and protein–protein interaction network analysis to investigate the signaling pathways and functions of the identified DEMs as well as the interactions between their target genes. Furthermore, we selected two upregulated and top 10 downregulated DEMs with the highest |log2FC| values as the key microRNAs, which may serve as potential biomarkers for MetS. RT-qPCR was performed to validated these result. Finally, hsa-miR-526b-5p, hsa-miR-6516-5p was identified as the novel biomarkers for MetS.View... Based on the high numbers of miRNAs and their involvement in virtually all biological processes, including immune regulation, multiple studies investigated their biomarker potential in the context of islet autoimmunity and T1D (42,56,66,(85)(86)(87)(88)(89)(90)(91). In contrast to the majority of disease symptoms, biomarkers enable the objectively quantifiable characterization of a disease and its progression. ...... In diabetic NOD mice, miR409-3p was reduced in the plasma as well as in islet infiltrates, and miR409-3p levels were associated with insulitis severity. In human patients with recent onset of T1D, plasma levels of miR409-3p were comparably reduced and correlated inversely with the levels of HbA1c (88). A recent work systematically reviewed and analyzed profiles of circulating miRNAs in T1D patients and suggested a combination of 11 miRNAs (miR21-5p, miR24-3p, miR100-5p, miR146a-5p, miR148a-3p, miR150-5p, miR181a-5p, miR210-5p, miR342-3p, miR375, miR1275), which are involved in several facets of immune regulation and beta cell function, as biomarkers for T1D (89). ...miRNA-Mediated Immune Regulation in Islet Autoimmunity and Type 1 DiabetesArticleFull-text availableNov 2020Martin G. SchermCarolin DanielThe important role of microRNAs as major modulators of various physiological processes, including immune regulation and homeostasis, has been increasingly recognized. Consequently, aberrant miRNA expression contributes to the defective regulation of T cell development, differentiation, and function. This can result in immune activation and impaired tolerance mechanisms, which exert a cardinal function for the onset of islet autoimmunity and the progression to T1D. The specific impact of miRNAs for immune regulation and how miRNAs and their downstream targets are involved in the pathogenesis of islet autoimmunity and T1D has been investigated recently. These studies revealed that increased expression of individual miRNAs is involved in several layers of tolerance impairments, such as inefficient Treg induction and Treg instability. The targeted modulation of miRNAs using specific inhibitors, resulting in improved immune homeostasis, as well as improved methods for the targeting of miRNAs, suggest that miRNAs, especially in T cells, are a promising target for the reestablishment of immune tolerance.View... A SOP was followed to collect plasma samples as previously described [41]. Blood was collected in BD Vacutainer K2-EDTA tubes (BD Biosciences, San Jose, CA, USA), inverted 5 times and stored upright at room temperature until ready for processing. ...MicroRNA Expression in the Aqueous Humor of Patients with Diabetic Macular EdemaArticleFull-text availableOct 2020INT J MOL SCI Giuseppina Emanuela Grieco Guido SebastianiChiara Maria EandiGiovanni NeriGian Marco TosiWe identified and compared secreted microRNA (miRNA) expression in aqueous humor (AH) and plasma samples among patients with: type 2 diabetes mellitus (T2D) complicated by non-proliferative diabetic retinopathy (DR) associated with diabetic macular edema (DME) (DME group: 12 patients); T2D patients without DR (D group: 8 patients); and non-diabetic patients (CTR group: 10 patients). Individual patient AH samples from five subjects in each group were profiled on TaqMan Low Density MicroRNA Array Cards. Differentially expressed miRNAs identified from profiling were then validated in single assay for all subjects. The miRNAs validated in AH were then evaluated in single assay in plasma. Gene Ontology (GO) analysis was conducted. From AH profiling, 119 mature miRNAs were detected: 86 in the DME group, 113 in the D group and 107 in the CTR group. miRNA underexpression in the DME group was confirmed in single assay for let-7c-5p, miR-200b-3p, miR-199a-3p and miR-365-3p. Of these four, miR-199a-3p and miR-365-3p were downregulated also in the plasma of the DME group. GO highlighted 54 validated target genes of miR-199a-3p, miR-200b-3p and miR-365-3p potentially implied in DME pathogenesis. Although more studies are needed, miR-200b-3p, let-7c-5p, miR-365-3p and miR-199a-3p represent interesting molecules in the study of DME pathogenesis.View... Beta cells also release intact granules [48] and exosomes carrying antigens such as proinsulin, GAD65 and islet antigen 2 (IA-2) [49], which are endocytosed more efficiently than soluble proteins/peptides and may further feed distant APCs. Beta cell-derived microRNAs released into the bloodstream, either in soluble form or associated with exosomes [50], may exert additional immunomodulatory effects. ...Presumption of innocence for beta cells: why are they vulnerable autoimmune targets in type 1 diabetes?ArticleFull-text availableOct 2020Diabetologia Roberto MalloneDecio L EizirikIt is increasingly appreciated that the pathogenic mechanisms of type 1 diabetes involve both the autoimmune aggressors and their beta cell targets, which engage in a conflicting dialogue within and possibly outside the pancreas. Indeed, autoimmune CD8⁺ T cells, which are the final mediators of beta cell destruction, circulate at similar frequencies in type 1 diabetic and healthy individuals. Hence a universal state of ‘benign’ islet autoimmunity exists, and we hypothesise that its progression to type 1 diabetes may at least partially rely on a higher vulnerability of beta cells, which play a key, active role in disease development and/or amplification. We posit that this autoimmune vulnerability is rooted in some features of beta cell biology: the stress imposed by the high rate of production of insulin and other granule proteins, their dense vascularisation and the secretion of their products directly into the bloodstream. Gene variants that may predispose individuals to this vulnerability have been identified, e.g. MDA5, TYK2, PTPN2. They interact with environmental cues, such as viral infections, that may drive this genetic potential towards exacerbated local inflammation and progressive beta cell loss. On top of this, beta cells set up compensatory responses, such as the unfolded protein response, that become deleterious in the long term. The relative contribution of immune and beta cell drivers may vary and phenotypic subtypes (endotypes) are likely to exist. This dual view argues for the use of circulating biomarkers of both autoimmunity and beta cell stress for disease staging, and for the implementation of both immunomodulatory and beta cell-protective therapeutic strategies.Graphical abstractView... C57Bl/6J, NOD-SCID, and NOD mice were housed and inbred in the animal facility of Katholieke Universiteit Leuven (KU Leuven, Leuven, Belgium) as previously described (27). All animal procedures were performed in accordance with the NIH guidelines for the care and use of laboratory animals and protocols were approved by the Ethics Committees of the KU Leuven. ...Pancreatic Alpha-Cells Contribute Together With Beta-Cells to CXCL10 Expression in Type 1 DiabetesArticleFull-text availableSep 2020 Laura Nigi Noemi Brusco Giuseppina E. Grieco Giada Licata Francesco DottaC-X-C Motif Chemokine Ligand 10 (CXCL10) is a pro-inflammatory chemokine specifically recognized by the ligand receptor CXCR3 which is mostly expressed in T-lymphocytes. Although CXCL10 expression and secretion have been widely associated to pancreatic islets both in non-obese diabetic (NOD) mice and in human type 1 diabetic (T1D) donors, the specific expression pattern among pancreatic endocrine cell subtypes has not been clarified yet. Therefore, the purpose of this study was to shed light on the pancreatic islet expression of CXCL10 in NOD, in C57Bl/6J and in NOD-SCID mice as well as in human T1D pancreata from new-onset T1D patients (DiViD study) compared to non-diabetic multiorgan donors from the INNODIA European Network for Pancreatic Organ Donors with Diabetes (EUnPOD). CXCL10 was expressed in pancreatic islets of normoglycaemic and new-onset diabetic NOD mice but not in C57Bl/6J and NOD-SCID mice. CXCL10 expression was increased in pancreatic islets of new-onset diabetic NOD mice compared to normoglycaemic NOD mice. In NOD mice, CXCL10 colocalized both with insulin and glucagon. Interestingly, CXCL10-glucagon colocalization rate was significantly increased in diabetic vs. normoglycaemic NOD mouse islets, indicating an increased expression of CXCL10 also in alpha-cells. CXCL10 was expressed in pancreatic islets of T1D patients but not in non-diabetic donors. The analysis of the expression pattern of CXCL10 in human T1D pancreata from DiViD study, revealed an increased colocalization rate with glucagon compared to insulin. Of note, CXCL10 was also expressed in alpha-cells residing in insulin-deficient islets (IDI), suggesting that CXCL10 expression in alpha cells is not driven by residual beta-cells and therefore may represent an independent phenomenon. In conclusion, we show that in T1D CXCL10 is expressed by alpha-cells both in NOD mice and in T1D patients, thus pointing to an additional novel role for alpha-cells in T1D pathogenesis and progression.View... Lower plasma levels of five miRNAs have been reported in NOD mice compared with normoglycaemic mice [29]. Among these miRNAs was miR-409−3p, the expression of which was also decreased in the immune islet infiltrates of NOD and is known to regulate expression of immune-and metabolism-associated transcripts. ...... Expression of miR-409−3p was associated with the intensity of insulitis severity and infiltration of CD8+ central memory T cells. Notably, decreased plasma quantities of miR-409−3p was also reported in newly diagnosed patients with T1D in correlation with HbA1c levels [29]. In a separate study, glucose-induced changes in expression of 125 miRNAs in human endothelial cells (ECs) has been reported. ...Emerging roles of non-coding RNAs in the pathogenesis of type 1 diabetes mellitusArticleFull-text availableSep 2020BIOMED PHARMACOTHER Mohammad Taheri Reyhane Eghtedarian Marcel E. Dinger Soudeh Ghafouri-FardType 1 diabetes mellitus (T1D) is a lifelong autoimmune disorder that is increasingly prevalent in populations worldwide. As well as affecting adults, T1D is one of the most prevalent chronic childhood disorders. Several lines of evidence point to dysregulation of both cellular and humoral immune responses in this disorder. Several genetic loci have been associated with risk of T1D, implying the presence of a complex multifactorial pattern of inheritance for this disorder. Moreover, recent studies have reported dysregulation of long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) in animal models of T1D or clinical samples. Several immune-related molecules and pathways such as NF-κB, PI3K/Akt/FOXO, JAK, MAPK, mTOR and STAT pathways are regulated by non-coding RNAs in the context of T1D. Improved understanding of the role of lncRNAs and miRNAs in the pathogenesis of T1D would facilitate design of preventive therapeutic modalities. In the current review, we summarize the results of animal and human studies that report dysregulation of these transcripts and their function in T1D.View... A series of studies have revealed the regulation of microRNA in aging, tumor progression, metabolic diseases, and inflammation (Klieser et al., 2019;Majidinia et al., 2019;Nasr et al., 2019;Zhao et al., 2019). In recent years, a variety of microRNAs (miR-409-3p, miR-98-5p, miR-16-5p, etc.) have been proved to participate in DM progression and its complications (Duan et al., 2019;Khan et al., 2019;Ventriglia et al., 2019). Furthermore, numerous microRNAs (miR-3197, miR-2116-5p, miR-152, miR-34a, etc.) are identified as specificity biomarkers, and they participate directly in DR progression (Fu and Ou, 2019;Ji et al., 2019;Thounaojam et al., 2019). ...MicroRNA-29b-3p Promotes Human Retinal Microvascular Endothelial Cell Apoptosis via Blocking SIRT1 in Diabetic RetinopathyArticleFull-text availableJan 2020 Yong Zeng Zekai Cui Jian Liu Jiansu ChenShibo TangBackground: Diabetic retinopathy (DR) is a main complication of diabetes mellitus (DM). Recent studies have implicated microRNAs in human retinal microvascular endothelial cell (HRMEC) dysfunction. In this study, we aim to investigate the apoptotic promotion of miR-29b-3p by blocking SIRT1 in HRMEC for DR situation.Method: Blood samples were obtained from DR patients and controls. Dual-luciferase reporter assay using HEK-293T cells was performed to show the direct interaction of miR-29b-3p and the 3 UTR of SIRT1. HRMECs were exposed to 5.5 mmol/L of glucose (normal control), 5.5 mmol/L of glucose and 24.5 mmol/L of mannitol (osmotic pressure control), 30 mmol/L of glucose [hyperglycemia (HG)], 150 μmol/L of CoCl2 (hypoxia), and 30 mmol/L of glucose plus 150 μmol/L of CoCl2 (HG-CoCl2). To identify the regulating relationship between miR-29b-3p and SIRT1, HRMECs were transfected with miR-29b-3p mimics/inhibitors or their negative controls. SRT1720 was used as a SIRT1 agonist. Cell viability was assessed with the cell counting kit-8 (CCK-8) assay, and apoptotic cells were stained by one-step terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay kit. Gene and protein expression were assayed by quantitative real-time reverse transcriptase-PCR (RT-qPCR) and western blotting separately.Result: MiR-29b-3p was upregulated to 3.2-fold, and SIRT1 protein was downregulated to 65% in DR patients. Dual-luciferase reporter assay showed the direct interaction of miR-29b-3p and SIRT1. HRMECs were identified as 95% positive for CD31 and von Willebrand factor (vWF). MiR-29b-3p and Bax/Bcl-2 ratio was upregulated, whereas SIRT1 was downregulated in HRMECs in the HG-CoCl2 condition. Decreased cell viability and upregulated apoptosis were also found in HRMECs of the HG-CoCl2 condition. Upregulated miR-29b-3p decreased the expression of SIRT1 and increased the ratio of Bax/Bcl-2, whereas downregulated miR-29b-3p increased the expression of SIRT1 protein and downregulated the ratio of Bax/Bcl-2. SRT1720 rescued miR-29b-3p-induced HRMEC apoptosis via upregulating the expression of SIRT1 protein.Conclusion: The dysregulation of miR-29b-3p/SIRT1 is a potential mechanism of HRMEC apoptosis in DR. MiR-29b-3p/SIRT1 may be a potential therapeutic target for DR.ViewMicroRNAs, Parkinson’s Disease, and Diabetes MellitusArticleFull-text availableMar 2021INT J MOL SCI Hsiuying WangParkinson’s disease (PD) is a neurodegenerative disorder that affects 1% of the population over the age of 60. Diabetes Mellitus (DM) is a metabolic disorder that affects approximately 25% of adults over the age of 60. Recent studies showed that DM increases the risk of developing PD. The link between DM and PD has been discussed in the literature in relation to different mechanisms including mitochondrial dysfunction, oxidative stress, and protein aggregation. In this paper, we review the common microRNA (miRNA) biomarkers of both diseases. miRNAs play an important role in cell differentiation, development, the regulation of the cell cycle, and apoptosis. They are also involved in the pathology of many diseases. miRNAs can mediate the insulin pathway and glucose absorption. miRNAs can also regulate PD-related genes. Therefore, exploring the common miRNA biomarkers of both PD and DM can shed a light on how these two diseases are correlated, and targeting miRNAs is a potential therapeutic opportunity for both diseases.ViewHsa_circ_0054633 mediates apoptosis and insulin secretion in human pancreatic β cells through miR-409-3p/caspase-8 axisArticleApr 2021Rui SunWanli XueJuzhen ZhaoBackground: CircRNAs are reported to be aberrantly expressed and perform biological functions in diverse processes. This study aimed to investigate the potential involvement of hsa_circ_0054633 in high glucose (HG)‑induced diabetic model and its potential mechanism.Methods: The expression of hsa_circ_0054633, miR-409-3p and caspase-8 was detected by real-time PCR and western blotting. Cell viability, apoptosis and the protein levels of apoptosis-related factors were revealed by CCK-8 colorimetry, flow cytometry and western blotting, respectively. Insulin secretion was determined by enzyme-linked immunosorbent assay (ELISA) and the measurement of insulin-related transcription factors. The target association between miR-409-3p and hsa_circ_0054633 or caspase-8 was confirmed by dual-luciferase reporter assays and biotin-based pulldown assay.Results: Hsa_circ_0054633 was highly expressed and the expression of miR-409-3p was downregulated in serum of DM patients and HG-treated human pancreatic β cell line NES2Y. Further investigation indicated that hsa_circ_0054633 suppression promoted cell proliferation, inhibited apoptosis and facilitated insulin secretion in HG-treated NES2Y cells. Mechanical analysis revealed that hsa_circ_0054633 regulated caspase-8 expression via sponging miR-409-3p. Rescue experiments demonstrated that miR-409-3p knockdown or caspase-8 overexpression reversed the effects of hsa_circ_0054633 in HG-stimulated NES2Y cells.Conclusion: Inhibition of hsa_circ_0054633 protected against HG-induced NES2Y cell apoptosis and impairment of insulin secretion by regulating miR-409-3p/caspase-8 axis.ViewShow moreGet access to 30 million figuresJoin ResearchGate to access over 30 million figures and 135+ million publications – all in one place.Join for freeAdvertisementJoin ResearchGate to find the people and research you need to help your work.20+ million members135+ million publications700k+ research projectsJoin for free orDiscover by subject areaRecruit researchersJoin for freeLoginEmail Tip: Most researchers use their institutional email address as their ResearchGate loginPasswordForgot password? Keep me logged inLog inorContinue with GoogleWelcome back! 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